A Typical Experiment
A typical directed evolution experiment involves three steps:
- Diversification: The gene encoding the protein of interest is mutated and/or recombined at random to create a large library of gene variants. Techniques commonly used in this step are error-prone PCR and DNA shuffling.
- Selection: The library is tested for the presence of mutants (variants) possessing the desired property using a screen or selection. Screens enable the researcher to identify and isolate high-performing mutants by hand, while selections automatically eliminate all nonfunctional mutants.
- Amplification: The variants identified in the selection or screen are replicated manyfold, enabling researchers to sequence their DNA in order to understand what mutations have occurred.
Together, these three steps are termed a "round" of directed evolution. Most experiments involve more than one round. In these experiments, the "winners" of the previous round are diversified in the next round to create a new library. At the end of the experiment, all evolved protein or RNA mutants are characterized using biochemical methods.
Read more about this topic: Directed Evolution
Famous quotes containing the words typical and/or experiment:
“A building is akin to dogma; it is insolent, like dogma. Whether or no it is permanent, it claims permanence, like a dogma. People ask why we have no typical architecture of the modern world, like impressionism in painting. Surely it is obviously because we have not enough dogmas; we cannot bear to see anything in the sky that is solid and enduring, anything in the sky that does not change like the clouds of the sky.”
—Gilbert Keith Chesterton (1874–1936)
“What constitutes a real, live human being is more of a mystery than ever these days, and men—each one of whom is a valuable, unique experiment on the part of nature—are shot down wholesale.”
—Hermann Hesse (1877–1962)